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Department of Protein Structure Institute
of Physiology AS CR, v.v.i. Videnska
1083 142
00 Praha 4 – Krc Czech
Republic |
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PROJECTS |
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OBSILOVA´s
group |
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We study molecular
mechanisms by which
protein function can be regulated. In particular, we are interested in the regulation of proteins involved in cellular signal transduction. Our main goal is
to provide mechanistic understanding of their regulations by studying relationships between their structure and function. |
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PROBLEMATICS: |
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The study of structure
and function of 14-3-3 proteins and their complexes. Our research team has been studying the 14-3-3 proteins which are highly conserved regulatory molecules found in all eukaryotes. First they have been
isolated from the bovine brain and their unusual name “14-3-3”, originates from their elution
and migration pattern on two-dimensional DEAE-cellulose chromatography and starch gel electrophoresis.
14-3-3 proteins have the ability of
binding the functionally different signal proteins, including kinases, fosfatases and transmembrane receptors by changing their function. Through the functional
modulation of a wide range of
binding partners, 14-3-3 proteins are involved in many processes, including cell cycle regulation, metabolism control, apoptosis, and control of gene transcription.More
than 300 proteins have been described
as binding partners till now. Main
goal of these projects is mechanistic
understanding of the 14-3-3 protein function in the regulation of selected 14-3-3 binding partners: for example the
interaction of 14-3-3
protein with forkhead transcription factor
FOXO4, with the regulatory domain of the tyrosine hydroxylase, interaction
with ASK1 kinase, with the regulator of G-protein signaling RGS3 and phosducin. Recently, we have
been studying two yeast isoforms of 14-3-3 protein (Bmh1 and Bmh2) and the interaction
of Bmh with neutral trehalase in yeast. |
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Specific trehalase activity of pNth1 WT and mutants upon the Bmh1 activation in vitro
Veisova et. al., Biochemical Journal 2012,
443:663-670. |
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Structure of human 14-3-3 protein (zeta isoform) with the modeled C-terminal segment
Veisova et. al., Biochemistry 2010, 49(18):3853-61. |
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Continuous distribution of sedimentation coefficients,
c(s), for Bmh1 and Nth1 alone
and in the complex
Veisova
et. al., Biochemical Journal 2012, 443:663-670. |
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METHODS |
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• Biochemical approach (recombinant protein expression, site-directed
mutagenesis,
enzyme kinetics) • Biophysical approach (fluorescence spectroscopy, analytical
ultracentrifugation, isotermal titration
calorimetry, surface plasmon resonance, H/D
exchange, mass spectrometry, protein structure modeling, X-ray crystallography, molecular dynamics
simulations). • Crystallography of selected
complexes → These
methods enable us to better understand the details how is
regulated the activity and function of protein-protein complexes. |
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SOLVED
STRUCTURES |
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DNA-binding
domain of forkhead transcription factor FOXO4 bound to the DNA
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RGS domain of RGS3 (regulator of G-protein signaling)
Rezabkova
et. al., Journal of Structural Biology 2010,
170, 451-461. |
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TEISINGER´s group |
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The study of the cytoplasmatic domains of TRP channels. Transient receptor potential
(TRP) channels are a wide
family of non-selective ion channels responsible for monovalent and divalent cation influx into the cells.
Members of this family are involved in many sensory processes
such as invertebrate vision and hearing,
mammalian temperature-, mechano- and chemo-sensation. The TRP channels discovered so far can be divided into
seven subfamilies according to their primary structure: TRPV, TRPC,
TRPA, TRPM, TRPP, TRPML and TRPN. All are predicted to have six transmembrane helices (S1–S6)
and a pore-forming loop between S5 and S6, with varying sizes of intracellular amino and carboxy termini, and are thought to form tetrameric assemblies. Both the N- and C-terminal intracellular domains are comprised of many different domains that are responsible for binding different
compounds that can regulate the channels. Our goal is to provide the
structural insight into the interactions of TRP channels with ATP, calmodulin and PIP. |
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